Anti-Nucleolin antibody [4E2] - ChIP Grade (ab13541)

Overview

  • Product name
    Anti-Nucleolin antibody [4E2] - ChIP Grade
    See all Nucleolin primary antibodies
  • Description
    Mouse monoclonal [4E2] to Nucleolin - ChIP Grade
  • Tested applications
    Suitable for: ICC/IF, IHC-P, WB, ICC, Flow Cyt, ChIPmore details
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse
  • Immunogen

    Human nucleolin protein from Raji cell extract.

  • Positive control
    • Raji cell extract.

Properties

Applications

Our Abpromise guarantee covers the use of ab13541 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use at an assay dependent concentration.
IHC-P Use a concentration of 1 - 10 µg/ml. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 100 kDa.
ICC Use a concentration of 1 - 10 µg/ml.
Flow Cyt Use a concentration of 5 - 10 µg/ml.

ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ChIP Use at an assay dependent concentration. PubMed: 19615975(recombinant nucleolin binding in the HPV16 genome).

Target

  • Function
    Nucleolin is the major nucleolar protein of growing eukaryotic cells. It is found associated with intranucleolar chromatin and pre-ribosomal particles. It induces chromatin decondensation by binding to histone H1. It is thought to play a role in pre-rRNA transcription and ribosome assembly. May play a role in the process of transcriptional elongation. Binds RNA oligonucleotides with 5'-UUAGGG-3' repeats more tightly than the telomeric single-stranded DNA 5'-TTAGGG-3' repeats.
  • Sequence similarities
    Contains 4 RRM (RNA recognition motif) domains.
  • Post-translational
    modifications
    Some glutamate residues are glycylated by TTLL8. This modification occurs exclusively on glutamate residues and results in a glycine chain on the gamma-carboxyl group.
  • Cellular localization
    Nucleus > nucleolus. Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
  • Information by UniProt
  • Database links
  • Alternative names
    • C23 antibody
    • FLJ45706 antibody
    • MS1116 antibody
    • NCL antibody
    • Nucl antibody
    • NUCL_HUMAN antibody
    • Nucleolin antibody
    • Protein C23 antibody
    see all

Anti-Nucleolin antibody [4E2] - ChIP Grade images

  • Immunocytochemistry/ Immunofluorescence analysis of 293 cells labeling Nucleolin with ab13541. Cells were fixed in chilled methanol for 30 min at −20°C, washed with PBS, and incubated in blocking solution (10% human serum in PBS) for 1 hour at room temperature. Cells were stained with Anti-Nucleolin antibody [4E2] - ChIP Grade (ab13541) diluted in blocking solution for 1 hour at room temperature in humidified chambers. Cells were washed with PBS, then incubated with secondary antibody diluted 1/200 in blocking solution for 1 hour at room temperature in opaque humidified chambers. Cells were washed with PBS, briefly rinsed in distilled H2O to remove salts, then mounted on glass slides.

  • ab13541 at 1µg/ml staining Nucleolin in human colon cancer tissue section by Immunohistochemistry (Formalin/ PFA fixed paraffin-embedded tissue sections).

  • All lanes : Anti-Nucleolin antibody [4E2] - ChIP Grade (ab13541) at 1 µg/ml

    Lane 1 : Cell lysates prepared from human HL60 cells
    Lane 2 : Cell lysates prepared from human ZR-75-1 cells


    Predicted band size : 100 kDa
  • Overlay histogram showing HeLa cells stained with ab13541 (red line). The cells were fixed with 100% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab13541, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • ab 13541 at a 1/2000 dilution staining U2OS cells by Immunocytochemistry. The antibody was incubated with the cells for 1 hour and then detected using a Alexa-Fluor ® 594 chicken anti-mouse antibody.

    This image is courtesy of an Abreview submitted by Raffaela Santoro on 16 March 2006.

    See Abreview

References for Anti-Nucleolin antibody [4E2] - ChIP Grade (ab13541)

This product has been referenced in:
  • Lun X  et al. Disulfiram when Combined with Copper Enhances the Therapeutic Effects of Temozolomide for the Treatment of Glioblastoma. Clin Cancer Res 22:3860-75 (2016). WB ; Human . Read more (PubMed: 27006494) »
  • Homma S  et al. Nuclear bodies reorganize during myogenesis in vitro and are differentially disrupted by expression of FSHD-associated DUX4. Skelet Muscle 6:42 (2016). ICC/IF ; Human . Read more (PubMed: 27906075) »

See all 29 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (ZR75-1 cells)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
20 µg
Specification
ZR75-1 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Apr 10 2017

Application
ChIP
Detection step
Real-time PCR
Sample
Human Cell lysate - whole cell (Breast)
Specification
Breast
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Username

Abcam user community

Verified customer

Submitted Jun 02 2014

Application
Western blot
Loading amount
1e+006 cells
Gel Running Conditions
Reduced Denaturing (12%)
Sample
Human Cell lysate - whole cell (Breast)
Specification
Breast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted May 27 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Human Tissue sections (Breast)
Specification
Breast
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted May 23 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (primary smooth muscle cells)
Loading amount
25 µg
Specification
primary smooth muscle cells
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Yiwen Liu

Verified customer

Submitted Mar 20 2012

Merci de nous avoir contactés. L'équipe qui produit l'anticorps ab13541 m'a informé que l'épitope n'avait malheureusement pas été déterminé. Désolé de ne pas pouvoir vous donner plus d'information sur ce produit.

Thank you for contacting us. The source of the antibody ab13541 informed me that, unfortunately, the epitope has not been mapped. The only information we have is that ab13541 is directed against the Human nucleotin, reference SwisProt # P19338 (http...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (glioma cell)
Specification
glioma cell
Fixative
Paraformaldehyde
Permeabilization
Yes - NP-40+BSA in PBS
Username

Ms. YOO MIRAN

Verified customer

Submitted May 14 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (prostate cancer cells)
Loading amount
20 µg
Specification
prostate cancer cells
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Sebastiano Battaglia

Verified customer

Submitted Dec 05 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (A431 Xenograft in mouse)
Specification
A431 Xenograft in mouse
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated
Blocking step
Dako Protein Block as blocking agent for 15 minute(s) · Concentration: pre dilute
Username

Abcam user community

Verified customer

Submitted Apr 27 2007

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